Methicillin-resistant Staphylococcus aureus (MRSA) is the most common multidrug-resistant pathogen in healthcare settings worldwide. The use of active detection and isolation (ADI) is a proven method to prevent MRSA infections that requires the detection of the MRSA-colonized patient through active surveillance testing (AST). Our long-term goal is to develop accurate tests with rapid turn-around times (<15 minutes) that could increase the efficacy of ADI and AST for MRSA prevention and control programs and molecular diagnosis across all healthcare settings. Our hypothesis is that droplet PCR will provide accurate test results with reduced analysis time, complexity and cost thereby making nucleic acid amplification tests viable near point-of- care. We propose to develop two in vitro point-of-care nucleic acid amplification tests for detecting MRSA in patient specimens. In addition to supporting development, this project would enable a mechanism and provide support for partnership between technology developers in private industry at QuantaLife Inc., an academic collaborator at Stanford University, and healthcare professionals at the University of Mississippi Medical Center (UMMC). QuantaLife scientists and engineers will develop prototype droplet PCR instrumentation and accompanying assays. UMMC has the requisite clinical expertise, specifically in the area of healthcare acquired infections, to guide test design and requirements and critically evaluate droplet PCR performance and potential impact in real-world clinical settings. The Stanford Microfluidics Laboratory is world-renowned as a leader in electrokinetic transport in microfluidic systems and has significant experience modeling and understanding physical processes at the nano- and micro-scale for biological sample preparation, and will support QuantaLife in development of automated DNA pre-processing modules. These partnerships could accelerate the translation of droplet PCR technology from the laboratory into an instrument for near-point-of- care testing thereby impacting healthcare sooner. PUBLIC HEALTH RELEVANCE: We propose to develop rapid tests for detecting the DNA of antibiotic resistant bacteria from patient samples. If implemented near the point of patient care these tests could quickly identify people who carry these bacteria and prevent subsequent infections within hospitals and clinics. This technology makes billions of copies of DNA in tiny droplets that accurately fingerprints the bacteria.